Inhibition of lncRNA NEAT1 protects endothelial cells against hypoxia/reoxygenation‑induced NLRP3 inflammasome activation by targeting the miR‑204/BRCC3 axis

Yao,Tao; Song,Yiting; Li,Shutao; Gu,Jing; Yan,Xuetao

doi:10.3892/mmr.2021.12548

Inhibition of lncRNA NEAT1 protects endothelial cells against hypoxia/reoxygenation‑induced NLRP3 inflammasome activation by targeting the miR‑204/BRCC3 axis

Authors:
- Tao Yao
- Yiting Song
- Shutao Li
- Jing Gu
- Xuetao Yan
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Affiliations: Department of Anesthesiology, Shenzhen Bao'an Maternity and Child Health Hospital, Shenzhen, Guangdong 518100, P.R. China
Published online on: November 29, 2021 https://doi.org/10.3892/mmr.2021.12548
Article Number: 32
Copyright: © Yao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cardiovascular ischemia/reperfusion (I/R) injury is primarily caused by oxygen recovery after prolonged hypoxia. Previous studies found that the long non coding RNA (lncRNA) nuclear enriched abundant transcript 1 (NEAT1) was involved in cardiovascular pathology, and that NOD‑like receptor protein 3 (NLRP3) inflammasome activation‑dependent pyroptosis played a key role in cardiovascular I/R injury. The present study aimed to explore the molecular mechanism of I/R pathogenesis in order to provide novel insights for potential future therapies. Cell viability and lactate dehydrogenase enzyme activity assays were used to detect cell injury after human umbilical vein endothelial cells (HUVECs) were subjected to hypoxia/reoxygenation (H/R). The expression of the NEAT1/microRNA (miR)‑204/BRCA1/BRCA2‑containing complex subunit 3 (BRCC3) axis was examined by reverse transcription‑quantitative PCR, and the associations among genes were confirmed by luciferase reporter assays. Western blotting and ELISA were used to measure the level of NLRP3 inflammasome activation‑dependent pyroptosis. The results demonstrated that NEAT1, BRCC3 expression and NLRP3 inflammasome activation‑dependent pyroptosis were significantly increased in H/R‑injured HUVECs, whereas silencing BRCC3 or NEAT1 attenuated H/R‑induced injury and pyroptosis. NEAT1 positively regulated BRCC3 expression via competitively binding with miR‑204. Moreover, NEAT1 overexpression counteracted miR‑204 mimic‑induced injury, BRCC3 expression and NLRP3 inflammasome activation‑dependent pyroptosis. Taken together, these findings demonstrated that inhibition of lncRNA NEAT1 protects HUVECs against H/R‑induced NLRP3 inflammasome activation by targeting the miR‑204/BRCC3 axis.

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1	Chen J, Jiang Z, Zhou X, Sun X, Cao J, Liu Y and Wang X: Dexmedetomidine Preconditioning Protects Cardiomyocytes Against Hypoxia/Reoxygenation-Induced Necroptosis by Inhibiting HMGB1-Mediated Inflammation. Cardiovasc Drugs Ther. 33:45–54. 2019. View Article : Google Scholar : PubMed/NCBI
2	Burzynski LC and Clarke MCH: Death Is Coming and the Clot Thickens, as Pyroptosis Feeds the Fire. Immunity. 50:1339–1341. 2019. View Article : Google Scholar : PubMed/NCBI
3	An N, Gao Y, Si Z, Zhang H, Wang L, Tian C, Yuan M, Yang X, Li X, Shang H, et al: Regulatory Mechanisms of the NLRP3 Inflammasome, a Novel Immune-Inflammatory Marker in Cardiovascular Diseases. Front Immunol. 10:15922019. View Article : Google Scholar : PubMed/NCBI
4	Py BF, Kim MS, Vakifahmetoglu-Norberg H and Yuan J: Deubiquitination of NLRP3 by BRCC3 critically regulates inflammasome activity. Mol Cell. 49:331–338. 2013. View Article : Google Scholar : PubMed/NCBI
5	Su Q, Liu Y, Lv XW, Ye ZL, Sun YH, Kong BH and Qin ZB: Inhibition of lncRNA TUG1 upregulates miR-142-3p to ameliorate myocardial injury during ischemia and reperfusion via targeting HMGB1- and Rac1-induced autophagy. J Mol Cell Cardiol. 133:12–25. 2019. View Article : Google Scholar : PubMed/NCBI
6	Liu H, Xu D, Zhong X, Xu D, Chen G, Ge J and Li H: lncRNA-mRNA competing endogenous RNA network depicts transcriptional regulation in ischaemia reperfusion injury. J Cell Mol Med. 23:2272–2276. 2019. View Article : Google Scholar : PubMed/NCBI
7	Du XJ, Wei J, Tian D, Yan C, Hu P, Wu X, Yang W and Hu X: NEAT1 promotes myocardial ischemia-reperfusion injury via activating the MAPK signaling pathway. J Cell Physiol. 234:18773–18780. 2019. View Article : Google Scholar : PubMed/NCBI
8	Wei Z, Qiao S, Zhao J, Liu Y, Li Q, Wei Z, Dai Q, Kang L and Xu B: miRNA-181a over-expression in mesenchymal stem cell-derived exosomes influenced inflammatory response after myocardial ischemia-reperfusion injury. Life Sci. 232:1166322019. View Article : Google Scholar : PubMed/NCBI
9	Luo M, Sun Q, Zhao H, Tao J and Yan D: Long noncoding RNA NEAT1 sponges miR-495-3p to enhance myocardial ischemia-reperfusion injury via MAPK6 activation. J Cell Physiol. 235:105–113. 2020. View Article : Google Scholar : PubMed/NCBI
10	Deng F, Wang S, Cai S, Hu Z, Xu R, Wang J, Feng D and Zhang L: Inhibition of Caveolae Contributes to Propofol Preconditioning-Suppressed Microvesicles Release and Cell Injury by Hypoxia-Reoxygenation. Oxid Med Cell Longev. 2017:35421492017. View Article : Google Scholar : PubMed/NCBI
11	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
12	Xue X and Luo L: lncRNA HIF1A-AS1 contributes to ventricular remodeling after myocardial ischemia/reperfusion injury by adsorption of microRNA-204 to regulating SOCS2 expression. Cell Cycle. 18:2465–2480. 2019. View Article : Google Scholar : PubMed/NCBI
13	Luo H, Wang J, Liu D, Zang S, Ma N, Zhao L, Zhang L, Zhang X and Qiao C: The lncRNA H19/miR-675 axis regulates myocardial ischemic and reperfusion injury by targeting PPARα. Mol Immunol. 105:46–54. 2019. View Article : Google Scholar : PubMed/NCBI
14	Zhu P, Yang M, Ren H, Shen G, Chen J, Zhang J, Liu J and Sun C: Long noncoding RNA MALAT1 downregulates cardiac transient outward potassium current by regulating miR-200c/HMGB1 pathway. J Cell Biochem. 119:10239–10249. 2018. View Article : Google Scholar : PubMed/NCBI
15	Jiang N, Xia J, Jiang B, Xu Y and Li Y: TUG1 alleviates hypoxia injury by targeting miR-124 in H9c2 cells. Biomed Pharmacother. 103:1669–1677. 2018. View Article : Google Scholar : PubMed/NCBI
16	Ren L, Chen S, Liu W, Hou P, Sun W and Yan H: Downregulation of long non-coding RNA nuclear enriched abundant transcript 1 promotes cell proliferation and inhibits cell apoptosis by targeting miR-193a in myocardial ischemia/reperfusion injury. BMC Cardiovasc Disord. 19:1922019. View Article : Google Scholar : PubMed/NCBI
17	Zhu Y, Yin X, Li J and Zhang L: Overexpression of microRNA-204-5p alleviates renal ischemia-reperfusion injury in mice through blockage of Fas/FasL pathway. Exp Cell Res. 381:208–214. 2019. View Article : Google Scholar : PubMed/NCBI
18	Yu SY, Dong B, Fang ZF, Hu XQ, Tang L and Zhou SH: Knockdown of lncRNA AK139328 alleviates myocardial ischaemia/reperfusion injury in diabetic mice via modulating miR-204-3p and inhibiting autophagy. J Cell Mol Med. 22:4886–4898. 2018. View Article : Google Scholar : PubMed/NCBI
19	Yan H, Liang H, Liu L, Chen D and Zhang Q: Long noncoding RNA NEAT1 sponges miR-125a-5p to suppress cardiomyocyte apoptosis via BCL2L12. Mol Med Rep. 19:4468–4474. 2019.PubMed/NCBI
20	Wang SM, Liu GQ, Xian HB, Si JL, Qi SX and Yu YP: lncRNA NEAT1 alleviates sepsis-induced myocardial injury by regulating the TLR2/NF-κB signaling pathway. Eur Rev Med Pharmacol Sci. 23:4898–4907. 2019.PubMed/NCBI
21	Yan L, Shi E, Jiang X, Shi J, Gao S and Liu H: Inhibition of MicroRNA-204 Conducts Neuroprotection Against Spinal Cord Ischemia. Ann Thorac Surg. 107:76–83. 2019. View Article : Google Scholar : PubMed/NCBI
22	Li JY, Yao YM and Tian YP: Ferroptosis: A Trigger of Proinflammatory State Progression to Immunogenicity in Necroinflammatory Disease. Front Immunol. 12:7011632021. View Article : Google Scholar : PubMed/NCBI
23	Deng F, Zhao BC, Yang X, Lin ZB, Sun QS, Wang YF, Yan ZZ, Liu WF, Li C, et al: The gut microbiota metabolite capsiate promotes Gpx4 expression by activating TRPV1 to inhibit intestinal ischemia reperfusion-induced ferroptosis. Gut Microbes. 13:1–21. 2021. View Article : Google Scholar
24	Ye L, He S, Mao X, Zhang Y, Cai Y and Li S: Effect of Hepatic Macrophage Polarization and Apoptosis on Liver Ischemia and Reperfusion Injury During Liver Transplantation. Front Immunol. 11:11932020. View Article : Google Scholar : PubMed/NCBI
25	Martin PK and Cadwell K: Regulation of interferon signaling in response to gut microbes by autophagy. Gut Microbes. 11:126–134. 2020. View Article : Google Scholar : PubMed/NCBI
26	Deng Y, Wu S, Yang Y, Meng M, Chen X, Chen S, Li L, Gao Y, Cai Y, Imani S, Chen B, Li S, Deng Y and Li X: Unique Phenotypes of Heart Resident Type 2 Innate Lymphoid Cells. Front Immunol. 11:8022020. View Article : Google Scholar : PubMed/NCBI